Apparatus and method for real-time measurement of cellular response

• US 6,379,917 B1
• Filed: 07/28/2000
• Issued: 04/30/2002
• Est. Priority Date: 08/02/1996
• Status: Expired due to Term

• Current Assignee: Caliper Life Sciences Incorporated (Revvity, Inc.)
• Original Assignee: AXIM Biotechnologies, Inc. (Axiomtek Co., Ltd.)
• Inventors: Okun, Alex, Okun, Ilya
• Primary Examiner(s): Leary, Louise N.
• Application Number:
  US09/627,642
• Time in Patent Office: 641 Days
• Field of Search: 435/29, 435/4, 435/7.1, 435/283.1, 422/50, 422/68.1, 364/148.05, 364/528.01
• US Class Current: 435/29
• CPC Class Codes: G01N 2510/00 Detection of programmed cell death, i.e. apoptosis
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/502 for testing non-proliferative effects
• CPC Class Codes: G01N 33/6872 Intracellular protein regulatory factors and their receptors, e.g. including ion channels
• CPC Class Codes: G01N 35/00 Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
• CPC Class Codes: G01N 35/1095 for supplying the samples to flow-through analysers for a specific analyser see relevant groups, e.g. under G01N15/00, G01N21/00, G01N27/00, G01N30/00, H01J49/00

Abstract

An apparatus and method for real-time measurement of a cellular response of a test compound or series of test compounds (303) on a flowing suspension of cells (349), in which a homogenous suspension of each member of a series of cell types (349) is combined with a concentration of a test compound (303), directed through a detection zone (355), and a cellular response of the living cells is measured in real time as the cells in the test mixture are flowing through the detection zone (355). The apparatus may be used in automated screening of libraries of compounds, and is capable of real-time variation of concentrations of test and standard compounds and generation of dose/response profiles within a short timespan.

Claims

1. An apparatus for measuring the concentration-dependent response of living cells to an agonist, comprising:

a cell suspension input that provides a constant flow of living cells;

an agonist input that provides an agonist;

a mixing zone, fluidly coupled to said cell suspension input and said agonist input, for mixing said agonist with said living cells;

a detector, fluidly coupled to the mixing zone, for measuring a response of the living cells to the agonist as said cells flow past said detector; and

a valve fluidly coupled to said detector for temporarily interrupting the flow of cells past the detector.

2. The apparatus of claim 1, wherein said cell suspension input further comprises a flow directing means having an intake port connected to said cell suspension and an output port fluidly connected to said mixing zone.

3. The apparatus of claim 2, wherein said flow directing means is a pump.

4. The apparatus of claim 1, wherein said agonist input comprises at least one reservoir for maintaining an agonist solution.

5. The apparatus of claim 1, wherein said agonist input comprises a multi-well plate for maintaining a plurality of agonist solutions.

6. The apparatus of claim 1, wherein said valve is controlled by a computer.

7. The apparatus of claim 1, wherein said mixing zone comprises a mixer that mixes said cell suspension with said agonist and a buffer.

8. The apparatus of claim 1, wherein said detector comprises an optical detector for measuring changes in the optical properties of said living cells.

9. The apparatus of claim 8, wherein said optical detector is a spectrophotometer.

10. The apparatus of claim 8, wherein said optical detector is a spectrofluorometer.

11. The apparatus of claim 8, wherein said optical detector is a flow cytometer.

12. A method for determining a response of living cells to an agonist, comprising:

providing a constant flow of an agonist;

providing a constant flow of living cells;

combining said flow of agonist with said flow of living cells to form a combined flow of said agonist and said living cells;

directing said combined flow of said agonist and said cells to a detector;

controlling the flow of said agonist and said cells through the detector by actuating a valve, wherein a response of the living cells to the varying concentrations of the agonist is measured as said cells are laced within said detector.

13. The method of claim 12, wherein combining said flow of agonist with said flow of living cells comprising introducing said agonist and said living cells flow into a mixing zone.

14. The method of claim 13, comprising determining a concentration-dependent response of said cells to said agonist after combining said agonist with said cells in said mixing zone.

15. The method of claim 14, wherein said response of said cells comprises a change in a spectral characteristic of said cells, induced by said agonist.

16. The method of claim 14, comprising determining a dose-response curve of said agonist on said living cells.

17. The method of claim 12, comprising using a computer to actuate said valve.